The isolation and identification of indigoid pigments from urine

Jackson, A. H.; Jenkins, R.T.; Grinstein, Moisés; Sancovich, A-M.Ferramola de; Sancovich, H. A.

doi:10.1016/0009-8981(88)90330-0

Cited by 16 publications

(8 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is just possible to directly observe the NMR spectrum of indigo [ 86 ], but the insolubility in the usual solvents at normal temperatures precludes this for dibromoindigo. The observation of the solid state NMR spectrum should be possible, but has not been reported.…”

Section: The Properties Of 66’-dibromoindigomentioning

confidence: 99%

Tyrian Purple: 6,6’-Dibromoindigo and Related Compounds

2001

View full text Add to dashboard Cite

show abstract

Section: The Properties Of 66’-dibromoindigomentioning

confidence: 99%

Tyrian Purple: 6,6’-Dibromoindigo and Related Compounds

2001

View full text Add to dashboard Cite

show abstract

“…Indirubin and indigo have been found in the urine of patients with acute myelomonocytic leukemia, porphyrinuria, and purple urine bag syndrome (19,20), but they have never been reported in urine from normal donors until now. We measured the concentrations of indirubin and indigo in urine that was processed within 15 min after excretion to minimize the potential effects of bacterial action (21).…”

Section: Indirubin As An Endogenous Ahr Ligand 31476mentioning

confidence: 99%

Indirubin and Indigo Are Potent Aryl Hydrocarbon Receptor Ligands Present in Human Urine

Adachi¹,

Mori²,

Matsui³

et al. 2001

Journal of Biological Chemistry

357

257

View full text Add to dashboard Cite

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates genes involved in xenobiotic metabolism, cellular proliferation, and differentiation. Numerous xenobiotic and biological compounds are known to interact with AhR, but it remains an orphan receptor, because its physiological ligand is unknown. We identified AhR ligands in human urine using a yeast AhR signaling assay and then characterized their properties. Two ligands, indirubin and indigo, were both present at average concentrations of ϳ0.2 nM in the urine of normal donors. Indirubin was also detected in fetal bovine serum and contributed half of the total AhR ligand activity. The activities of indirubin and indigo were comparable with or more potent than that of the archetypal ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin, in yeast AhR activation assays. We suggest that the endogenous levels and potencies of indirubin and indigo are such that they activate AhR-mediated signaling mechanisms in vivo. AhR,1 also called the dioxin receptor, is a ligand-activated transcription factor that is present in most cell and tissue types of the body (1). AhR-mediated signaling is required for potent xenobiotic ligands such as TCDD and polychlorinated biphenyls to produce toxic responses (2, 3). Toxic effects that are linked to xenobiotic AhR ligand exposures in animals include cancers, reproductive impairment, endometriosis, birth defects, and immunological impairment (4 -6). The toxic potential of xenobiotic AhR ligands is currently a major concern for regulatory agencies that are responsible for protecting public and environmental health. Although numerous xenobiotic ligands for AhR have been identified, the AhR is considered to be an orphan receptor, because its physiological ligand(s) and its function are not known. Tryptophan and other indole-containing compounds (7-9), bilirubin (10), 7-ketocholesterol (11), lipoxin A4 (12), flavones, and related compounds (13) interact with AhR to produce activation or inhibition of signal transduction. In general, the low levels, lack of potency, and restricted distribution of these compounds make them unlikely candidates as major regulators of AhR signaling in most tissue types. We reasoned that human urine might be a good place to search for endogenous AhR ligands, and we developed a methodology to detect and isolate such compounds. EXPERIMENTAL PROCEDURESMaterials-Blue rayon was kindly provided by Dr. Hayatsu (Okayama University, Okayama City, Japan). General chemicals, essentially analytical grade, were purchased from Wako (Kyoto, Japan). TCDD was purchased from CIL (Andover, MA, USA). Indigo and ␤-glucuronidase were purchased from Sigma. Indirubin was synthesized as described in Hoessel et al. (14). Indigo and indirubin were further purified by HPLC before use.Yeast Assay for AhR Ligand Activity-The assay procedure was essentially as described by Miller (15). The yeast strain YCM3 was grown overnight at 30°C in synthetic glucose medium lacking tryptophan. Test chemicals (dissolved in Me 2 SO...

show abstract

“…The chloroform layer was dried with anhydrous sodium sulfate and then removed in the rotary evaporator under reduced pressure. Chloroformextracted products were analyzed by thin-layer chromatography on activated silica gel plates (EM Science, Gibbstown, N.J.), which were developed with either toluene-acetone (4:1) (solvent A) or chloroform-acetone (97:3) (solvent B) (24). Incubations of strains mt-2 and F1 with indole-2-carboxylate and indole-3-carboxylate gave similar mixtures of three reaction products.…”

mentioning

confidence: 99%

Formation of indigo and related compounds from indolecarboxylic acids by aromatic acid-degrading bacteria: chromogenic reactions for cloning genes encoding dioxygenases that act on aromatic acids

Eaton

Chapman

1995

J Bacteriol

View full text Add to dashboard Cite

The p-cumate-degrading strain Pseudomonas putida F1 and the m-and p-toluate-degrading strain P. putida mt-2 transform indole-2-carboxylate and indole-3-carboxylate to colored products identified here as indigo, indirubin, and isatin. A mechanism by which these products could be formed spontaneously following dioxygenase-catalyzed dihydroxylation of the indolecarboxylates is proposed. Indolecarboxylates were employed as chromogenic substrates for identifying recombinant bacteria carrying genes encoding p-cumate dioxygenase and toluate dioxygenase. Dioxygenase gene-carrying bacteria could be readily distinguished as dark green-blue colonies among other colorless recombinant Escherichia coli colonies on selective agar plates containing either indole-2-carboxylate or indole-3-carboxylate.Dioxygenases are well recognized as important bacterial enzymes initiating aerobic catabolism of aromatic hydrocarbons (20,37). The actions of these enzymes furnish vicinal cis-dihydrodiols, precursors of o-diphenols which serve as substrates for ring cleavage enzymes. Of the many aromatic hydrocarbons shown to undergo reactions of this type, examples of a range of mono-and multinuclear substrates can be cited. Thus, benzene, toluene, ethylbenzene, cumene (isopropylbenzene), naphthalene, biphenyl, and phenanthrene have all been shown to yield cisdihydrodiols, in many instances with isotopic evidence of incorporation of both atoms of molecular oxygen. A useful chromogenic property of several of these enzyme systems is the ability to oxygenate the heterocyclic ring of indole, facilitating indigo formation (17). This property has been valuable in the cloning of aromatic hydrocarbon dioxygenase genes (10,11,16,29,30,45) and in the development of processes for the industrial production of indigo (32).Dioxygenase reactions leading to 1,2-diphenol formation and ring cleavage are also employed by bacteria in the catabolism of a number of aromatic carboxylic acids. This is the case for compounds such as benzoic acid (35, 36), phthalic acid (2, 15, 38), phenylpropionic (hydrocinnamic) acid (4, 42), m-and p-toluic acids (1, 43), and p-cumic acid (8). These are utilizable carbon sources in their own right but are also established intermediates in the degradation of, respectively, mandelic acid (18), phenanthrene (27, 28), 1-phenylalkanes with oddcarbon-number side chains (39), m-and p-xylene (1, 44), and p-cymene (8).To learn more about p-cumic acid, 2,3-dioxygenase, a study of the range of carboxylic acid substrates acted on by cells of the p-cymene-utilizing strain Pseudomonas putida F1 was undertaken. In the course of this work, it was noted that blue reaction products, reminiscent of indigo, were formed from certain indolecarboxylate isomers. Similar reactions were subsequently found with cells of P. putida mt-2 induced for m-and p-toluate 1,2-dioxygenase activity. The results reported here show that indigo and related products are, in fact, formed from indole-2-carboxylic acid and indole-3-carboxylic acid, and a reaction scheme is proposed...

show abstract

The isolation and identification of indigoid pigments from urine

Cited by 16 publications

References 14 publications

Tyrian Purple: 6,6’-Dibromoindigo and Related Compounds

Tyrian Purple: 6,6’-Dibromoindigo and Related Compounds

Indirubin and Indigo Are Potent Aryl Hydrocarbon Receptor Ligands Present in Human Urine

Formation of indigo and related compounds from indolecarboxylic acids by aromatic acid-degrading bacteria: chromogenic reactions for cloning genes encoding dioxygenases that act on aromatic acids

Contact Info

Product

Resources

About